title: Carbonic anhydrase activity from the gilthead sea bream (Sparus aurata) liver: the toxicological effects of heavy metals.
abstract: Many studies have shown that metal ions may lead to oxidative stress in biological systems. Accordingly, DNA damage, protein modification, enzyme inhibition and activation, lipid peroxidation and many other effects may occur in living organisms. Many different formations of metal ions may enter human cells along with water, air, and various foods, and humans are negatively affected by these conditions, either directly or indirectly. These effects may cause irreversible damage to human metabolism. In this study, the toxicological effects of heavy metals on carbonic anhydrase enzyme activity from the gilthead sea bream liver were investigated. The carbonic anhydrase enzyme was purified via affinity chromatography and had a specific activity of 6775.5EUmg(-1). The kinetics and characteristic properties, such as optimum pH, stable pH, optimum temperature, activation energy (Ea), activation enthalpy (ΔH), Q10, Km, and Vmax, were determined for the purified enzyme SDS-polyacrylamide gel electrophoresis showed a single band and molecular weight of the subunit was approximately 25kDa. Cd(II), Cu(II), Ni(II) and Ag(I) inhibited the enzyme activity in vitro. The type of inhibition and Ki values for these metals were calculated from Lineweaver-Burk plots as 17.74mM, 36.20mM, 12.85mM and 0.025mM for Cd(II), Cu(II), Ni(II) and Ag(I), respectively. All the metals were noncompetitive inhibitors.
authors.affiliation: Iğdır University, Faculty of Engineering, Department of Food Engineering, Iğdır, Turkey.
authors.forename: Elif Duygu
authors.initials: ED
authors.lastname: Kaya
authors.affiliation: Bayburt Univesity, Education Faculty, Department of Primary Education, Bayburt, Turkey. Electronic address: hsoyut@bayburt.edu.tr.
authors.forename: Hakan
authors.initials: H
authors.lastname: Söyüt
authors.affiliation: Atatürk University, Faculty of Sciences, Department of Chemistry, Erzurum, Turkey. Electronic address: beydemir@atauni.edu.tr.
authors.forename: Şükrü
authors.initials: Ş
authors.lastname: Beydemir
mesh_terms: D000818:Animals; D002257:Carbonic Anhydrase Inhibitors / Q000633:toxicity*; D002256:Carbonic Anhydrases / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004305:Dose-Response Relationship, Drug; D029941:Fish Proteins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000187:drug effects* / Q000201:enzymology; D019216:Metals, Heavy / Q000633:toxicity*; D008954:Models, Biological; D021541:Sea Bream / Q000378:metabolism*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D029941:Fish Proteins; D019216:Metals, Heavy; D014874:Water Pollutants, Chemical; D002256:Carbonic Anhydrases
keywords: CA; Carbonic anhydrase; Fish; Heavy metals; Inhibition; Liver; MW; RT; SDS-PAGE; carbonic anhydrase; molecular weight; rainbow trout; sodium dodecyl sulfate polyacrylamide gel electrophoresis